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Escherichia coli K-12 substr. MG1655 Enzyme: acetyl esterase



Gene: aes Accession Numbers: EG11101 (EcoCyc), b0476, ECK0470

Synonyms: ybaC

Regulation Summary Diagram: ?

Subunit composition of acetyl esterase = [Aes]2
         acetyl esterase = Aes

Summary:
Aes catalyzes hydrolysis of p-nitrophenyl esters of fatty acids, preferring substrates with short (<8 in length) acyl chains [Kanaya98]. Aes binds to the maltose operon regulator, MalT, and inhibits MalT transcriptional activation activity in competition with its inducer, maltotriose [Joly02]. Residues in MalT that are important for binding of Aes have been identified [Schlegel02a], and structure-based alanine scanning mutagenesis has identified residues in Aes that are responsible for interaction with MalT [Schiefner13]. Aes also binds to α-galactosidase; this interaction stimulates Aes esterase activity and inhibits α-galactosidase, suggesting that Aes may have a physiological role in regulation of metabolism of carbohydrates [Mandrich02]. Other proteins that interact with Aes have been identified [DAmbrosio08].

Aes has similarity to lipases and esterases [Peist97] and belongs to the hormone-sensitive lipase (HSL) protein family [Kanaya98]. The enzyme may be either monomeric [Kanaya98] or homodimeric [Schiefner13]; it has been crystallized [Sorrentino03, Gerber04], and crystal structures have been solved [Schiefner13]. Denaturant-induced unfolding of the enzyme has been studied [Del04], and a mutant with higher thermostability has been isolated [Farias07]. The catalytic triad was predicted to comprise Ser165, Asp262, and His292 [Kanaya98]; site-directed mutagenesis confirmed that these residues are required for catalytic activity. In addition, Asp164 is thought to be structurally important [Haruki99]. Point mutations in L97 and L209 increase enzymatic activity by increasing kcat and decreasing Km, respectively [Kobayashi12].

Overexpression of aes results in increased p-nitrophenyl acetate hydrolysis, compared to wild type, and this strain shows growth on triacetyl glycerol and repression of the maltose transport system [Peist97]. A genetic interaction between malT and aes has been observed [Peist97]. Mutations in MalT have differential effects on the sensitivity of MalT to Aes and MalY, which inhibit the activator in similar, yet distinct, ways [Joly02, Schlegel02a].

Aes is identical to esterase B, a protein marker used to distinguish E. coli phylogenetic groups [Lescat09].

Aes: "acetyl esterase" [Peist97]

Citations: [Del03]

Locations: cytosol

Map Position: [498,238 <- 499,197] (10.74 centisomes)
Length: 960 bp / 319 aa

Molecular Weight of Polypeptide: 36.034 kD (from nucleotide sequence), 37.0 kD (experimental) [Peist97 ]

Unification Links: ASAP:ABE-0001651 , DIP:DIP-9062N , EchoBASE:EB1093 , EcoGene:EG11101 , EcoliWiki:b0476 , ModBase:P23872 , OU-Microarray:b0476 , PortEco:aes , PR:PRO_000022064 , Protein Model Portal:P23872 , RefSeq:NP_415009 , RegulonDB:EG11101 , SMR:P23872 , String:511145.b0476 , UniProt:P23872

Relationship Links: InterPro:IN-FAMILY:IPR002168 , InterPro:IN-FAMILY:IPR013094 , InterPro:IN-FAMILY:IPR023508 , PDB:Structure:4KRX , PDB:Structure:4KRY , Pfam:IN-FAMILY:PF07859 , Prosite:IN-FAMILY:PS01173 , Prosite:IN-FAMILY:PS01174

Gene-Reaction Schematic: ?

GO Terms:

Biological Process: GO:0043433 - negative regulation of sequence-specific DNA binding transcription factor activity Inferred from experiment [Joly02]
GO:0051346 - negative regulation of hydrolase activity Inferred from experiment [Mandrich02]
GO:0008152 - metabolic process Inferred by computational analysis [GOA01a]
Molecular Function: GO:0005515 - protein binding Inferred from experiment [Joly02, Mandrich02]
GO:0008126 - acetylesterase activity Inferred from experiment [Kanaya98]
GO:0016787 - hydrolase activity Inferred from experiment Inferred by computational analysis [UniProtGOA11a, GOA01a, Kanaya98]
GO:0034338 - short-chain carboxylesterase activity Inferred from experiment [Kanaya98]
GO:0042803 - protein homodimerization activity Inferred from experiment [Schiefner13]
GO:0052689 - carboxylic ester hydrolase activity Inferred by computational analysis [UniProtGOA11a, GOA06, GOA01a]
Cellular Component: GO:0005737 - cytoplasm Author statement Inferred by computational analysis [UniProtGOA11, UniProtGOA11a, GOA06, GOA01a, Kanaya98]
GO:0005829 - cytosol Inferred by computational analysis [DiazMejia09]

MultiFun Terms: metabolism carbon utilization carbon compounds

Essentiality data for aes knockouts: ?

Growth Medium Growth? T (°C) O2 pH Osm/L Growth Observations
LB enriched Yes 37 Aerobic 6.95   Yes [Gerdes03, Comment 1]
LB Lennox Yes 37 Aerobic 7   Yes [Baba06, Comment 2]
M9 medium with 1% glycerol Yes 37 Aerobic 7.2 0.35 Yes [Joyce06, Comment 3]
MOPS medium with 0.4% glucose Yes 37 Aerobic 7.2 0.22 Yes [Baba06, Comment 2]

Credits:
Created 20-Aug-2013 by Keseler I , SRI International
Last-Curated ? 20-Aug-2013 by Keseler I , SRI International


Enzymatic reaction of: acetyl esterase

Synonyms: acetyl-esterase, acetylesterase

EC Number: 3.1.1.6

an acetic ester + H2O <=> an alcohol + acetate + H+

The reaction direction shown, that is, A + B ↔ C + D versus C + D ↔ A + B, is in accordance with the direction of enzyme catalysis.

The reaction is physiologically favored in the direction shown.

Alternative Substrates for an acetic ester: p-nitrophenyl butyrate [Kobayashi12 , Kanaya98 ] , p-nitrophenyl acetate [Khersonsky11 , Peist97 ]

Summary:
Kinetic parameters for various p-nitrophenyl esters of fatty acids have been measured [Kanaya98].

Note that acetyl-esterase is unstable at its optimal temperature [Farias07].

Kinetic Parameters:

Substrate
Km (μM)
kcat (sec-1)
kcat/Km (sec-1 μM-1)
Citations
p-nitrophenyl butyrate
170.0
29.0
[Kobayashi12]

T(opt): 65 °C [Farias07]


Sequence Features

Feature Class Location Citations Comment
Sequence-Conflict 18
[Mori90a, UniProt10]
Alternate sequence: K → N; UniProt: (in Ref. 8);
Protein-Segment 91 -> 93
[UniProt14]
UniProt: Involved in the stabilization of the negatively charged intermediate by the formation of the oxyanion hole; Sequence Annotation Type: short sequence motif; Non-Experimental Qualifier: by similarity.
Mutagenesis-Variant 103
[Haruki99, UniProt11]
Alternate sequence: H → A; UniProt: Reduces enzymatic efficiency.
Mutagenesis-Variant 128
[Haruki99, UniProt11]
Alternate sequence: E → A; UniProt: Reduces enzymatic efficiency.
Mutagenesis-Variant 163
[Haruki99, UniProt11]
Alternate sequence: G → A; UniProt: Diminishes catalytic efficiency.
Mutagenesis-Variant 164
[Haruki99, UniProt11]
Alternate sequence: D → A; UniProt: Strongly reduces enzymatic activity.
Mutagenesis-Variant 165
[Haruki99, UniProt11]
Alternate sequence: S → A; UniProt: Abolishes enzymatic activity.
Active-Site 165
[UniProt10a]
UniProt: Non-Experimental Qualifier: probable;
Mutagenesis-Variant 167
[Haruki99, UniProt11]
Alternate sequence: G → A; UniProt: Diminishes substrate affinity.
Mutagenesis-Variant 262
[Haruki99, UniProt11]
Alternate sequence: D → A; UniProt: Strongly reduces enzymatic activity.
Active-Site 262
[UniProt10a]
UniProt: Non-Experimental Qualifier: probable;
Mutagenesis-Variant 266
[Haruki99, UniProt11]
Alternate sequence: D → A; UniProt: Reduces enzymatic efficiency.
Sequence-Conflict 275 -> 319
[Miyamoto91, UniProt10]
Alternate sequence: LAAHQQPCEFKLYPGTLHAFLHYSRMMKTADEALRDGAQFFTAQL → VSCASAAL; UniProt: (in Ref. 2);
Mutagenesis-Variant 292
[Haruki99, UniProt11]
Alternate sequence: H → A; UniProt: Abolishes enzymatic activity.
Active-Site 292
[UniProt10a]
UniProt: Non-Experimental Qualifier: probable;


Gene Local Context (not to scale): ?

Transcription Unit:

Notes:

History:
10/20/97 Gene b0476 from Blattner lab Genbank (v. M52) entry merged into EcoCyc gene EG11101; confirmed by SwissProt match.


References

Baba06: Baba T, Ara T, Hasegawa M, Takai Y, Okumura Y, Baba M, Datsenko KA, Tomita M, Wanner BL, Mori H (2006). "Construction of Escherichia coli K-12 in-frame, single-gene knockout mutants: the Keio collection." Mol Syst Biol 2;2006.0008. PMID: 16738554

DAmbrosio08: D'Ambrosio C, Mandrich L, Rossi M, Scaloni A, Manco G (2008). "A proteomic approach to study Escherichia coli. Acetyl esterase interactors unveil a sequence motif involved in protein-protein interaction." Protein Pept Lett 15(4);333-40. PMID: 18473943

Del03: Del Vecchio P, Graziano G, Granata V, Barone G, Mandrich L, Rossi M, Manco G (2003). "Effect of trifluoroethanol on the conformational stability of a hyperthermophilic esterase: a CD study." Biophys Chem 104(2);407-15. PMID: 12878309

Del04: Del Vecchio P, Graziano G, Granata V, Farias T, Barone G, Mandrich L, Rossi M, Manco G (2004). "Denaturant-induced unfolding of the acetyl-esterase from Escherichia coli." Biochemistry 43(46);14637-43. PMID: 15544334

DiazMejia09: Diaz-Mejia JJ, Babu M, Emili A (2009). "Computational and experimental approaches to chart the Escherichia coli cell-envelope-associated proteome and interactome." FEMS Microbiol Rev 33(1);66-97. PMID: 19054114

Farias07: Farias T, Mandrich L, Rossi M, Manco G (2007). "Biochemical and thermostability features of acetyl esterase aes from Escherichia coli." Protein Pept Lett 14(2);165-9. PMID: 17305603

Gerber04: Gerber K, Schiefner A, Seige P, Diederichs K, Boos W, Welte W (2004). "Crystallization and preliminary X-ray analysis of Aes, an acetyl-esterase from Escherichia coli." Acta Crystallogr D Biol Crystallogr 60(Pt 3);531-3. PMID: 14993683

Gerdes03: Gerdes SY, Scholle MD, Campbell JW, Balazsi G, Ravasz E, Daugherty MD, Somera AL, Kyrpides NC, Anderson I, Gelfand MS, Bhattacharya A, Kapatral V, D'Souza M, Baev MV, Grechkin Y, Mseeh F, Fonstein MY, Overbeek R, Barabasi AL, Oltvai ZN, Osterman AL (2003). "Experimental determination and system level analysis of essential genes in Escherichia coli MG1655." J Bacteriol 185(19);5673-84. PMID: 13129938

GOA01a: GOA, DDB, FB, MGI, ZFIN (2001). "Gene Ontology annotation through association of InterPro records with GO terms."

GOA06: GOA, SIB (2006). "Electronic Gene Ontology annotations created by transferring manual GO annotations between orthologous microbial proteins."

Haruki99: Haruki M, Oohashi Y, Mizuguchi S, Matsuo Y, Morikawa M, Kanaya S (1999). "Identification of catalytically essential residues in Escherichia coli esterase by site-directed mutagenesis." FEBS Lett 454(3);262-6. PMID: 10431819

Joly02: Joly N, Danot O, Schlegel A, Boos W, Richet E (2002). "The Aes protein directly controls the activity of MalT, the central transcriptional activator of the Escherichia coli maltose regulon." J Biol Chem 277(19);16606-13. PMID: 11867639

Joyce06: Joyce AR, Reed JL, White A, Edwards R, Osterman A, Baba T, Mori H, Lesely SA, Palsson BO, Agarwalla S (2006). "Experimental and computational assessment of conditionally essential genes in Escherichia coli." J Bacteriol 188(23);8259-71. PMID: 17012394

Kanaya98: Kanaya S, Koyanagi T, Kanaya E (1998). "An esterase from Escherichia coli with a sequence similarity to hormone-sensitive lipase." Biochem J 332 ( Pt 1);75-80. PMID: 9576853

Khersonsky11: Khersonsky O, Malitsky S, Rogachev I, Tawfik DS (2011). "Role of chemistry versus substrate binding in recruiting promiscuous enzyme functions." Biochemistry 50(13);2683-90. PMID: 21332126

Kobayashi12: Kobayashi R, Hirano N, Kanaya S, Haruki M (2012). "Enhancement of the enzymatic activity of Escherichia coli acetyl esterase by a double mutation obtained by random mutagenesis." Biosci Biotechnol Biochem 76(11);2082-8. PMID: 23132590

Lescat09: Lescat M, Hoede C, Clermont O, Garry L, Darlu P, Tuffery P, Denamur E, Picard B (2009). "aes, the gene encoding the esterase B in Escherichia coli, is a powerful phylogenetic marker of the species." BMC Microbiol 9;273. PMID: 20040078

Mandrich02: Mandrich L, Caputo E, Martin BM, Rossi M, Manco G (2002). "The Aes protein and the monomeric alpha-galactosidase from Escherichia coli form a non-covalent complex. Implications for the regulation of carbohydrate metabolism." J Biol Chem 277(50);48241-7. PMID: 12374803

Miyamoto91: Miyamoto K, Nakahigashi K, Nishimura K, Inokuchi H (1991). "Isolation and characterization of visible light-sensitive mutants of Escherichia coli K12." J Mol Biol 219(3);393-8. PMID: 2051480

Mori90a: Mori H., Iida A., Teshiba S., Fujio T. (1990). Data submission to EMBL/GenBank/DDBJ databases on 1990-12.

Peist97: Peist R, Koch A, Bolek P, Sewitz S, Kolbus T, Boos W (1997). "Characterization of the aes gene of Escherichia coli encoding an enzyme with esterase activity." J Bacteriol 179(24);7679-86. PMID: 9401025

Schiefner13: Schiefner A, Gerber K, Brosig A, Boos W (2013). "Structural and mutational analyses of Aes, an inhibitor of MalT in Escherichia coli." Proteins. PMID: 23934774

Schlegel02a: Schlegel A, Danot O, Richet E, Ferenci T, Boos W (2002). "The N terminus of the Escherichia coli transcription activator MalT is the domain of interaction with MalY." J Bacteriol 184(11);3069-77. PMID: 12003949

Sorrentino03: Sorrentino N, De Simone G, Menchise V, Mandrich L, Rossi M, Manco G, Pedone C (2003). "Crystallization and preliminary X-ray diffraction studies of Aes acetyl-esterase from Escherichia coli." Acta Crystallogr D Biol Crystallogr 59(Pt 10);1846-8. PMID: 14501134

UniProt10: UniProt Consortium (2010). "UniProt version 2010-11 released on 2010-11-02 00:00:00." Database.

UniProt10a: UniProt Consortium (2010). "UniProt version 2010-07 released on 2010-06-15 00:00:00." Database.

UniProt11: UniProt Consortium (2011). "UniProt version 2011-06 released on 2011-06-30 00:00:00." Database.

UniProt14: UniProt Consortium (2014). "UniProt version 2014-01 released on 2014-01-01 00:00:00." Database.

UniProtGOA11: UniProt-GOA (2011). "Gene Ontology annotation based on the manual assignment of UniProtKB Subcellular Location terms in UniProtKB/Swiss-Prot entries."

UniProtGOA11a: UniProt-GOA (2011). "Gene Ontology annotation based on manual assignment of UniProtKB keywords in UniProtKB/Swiss-Prot entries."


Report Errors or Provide Feedback
Please cite the following article in publications resulting from the use of EcoCyc: Nucleic Acids Research 41:D605-12 2013
Page generated by SRI International Pathway Tools version 18.5 on Thu Nov 27, 2014, biocyc13.